PCR-SSCP and Sequencing of CXCR2 Receptor Gene in Vrindavani Cattle

Genetic markers associated with inflammatory responses during mastitis could aid in the selection of diseased cattle. One potential marker is CXCR2, a chemokine receptor required for neutrophil migration to infection sites. The objective of this experiment was to identify genetic polymorphism of CXCR2 gene and associate it with subclinical and clinical mastitis. Ninety five Vrindavani crossbred cows (42-mastitis tolerant and 53-clinical mastitis) that completed at least two full lactations were taken for study. Blood of selected crossbred cows was collected, and genomic DNA was isolated by phenol chloroform method. The DNA of good quality having OD ratio (260/280 nm) between 1.7-1.9 were used for further analysis. PCR-SSCP technique was used to reveal the polymorphism in 269bp fragments of CXCR2 gene. The 269 bp fragment of CXCR2 gene was found to be monomorphic in all the DNA samples of crossbred cows

Effect of heat shock protein 70 polymorphism on thermotolerance in Tharparkar cattle

Abstract Aim: Out of various members of heat shock protein (HSP) superfamily which act a molecular chaperon by binding to the denaturing protein thus stabilizing them and preserving their activity, HSP70 are of major importance in thermotolerance development. Thus, present investigation aimed at a screening of HSP70 gene for polymorphisms and possible differences in thermotolerance in Tharparkar breed of cattle. Materials and Methods: A 295 bp fragment of HSP70 gene was subjected to polymerase chain reaction-singlestrand conformation polymorphism (SSCP) followed by sequencing of different SSCP patterns in 64 Tharparkar cattle. A comparative thermotolerance of identified genotypes was analyzed using heat tolerance coefficients (HTCs) of animals for different seasons. Results: Three SSCP patterns and consequently two alleles namely A and B were documented in one fragment of HSP70 gene. On sequencing, one single-nucleotide polymorphism with G > T substitution was found at a position that led to a change of amino acid aspartate to tyrosine in allele A. It was found that in maintaining near normal average rectal temperature, genotype AA was superior (p≤0.01). Genotype AA, thus, was found to be most thermotolerant genotype with the highest HTC (p≤0.01). Conclusion: The polymorphism at HSP70 is expected to be a potent determinant for heat tolerance in cattle, which may aid in selection for thermotolerance in cattle

PCR-SSCP and Sequencing of CXCR2 Receptor Gene in Vrindavani Cattle

Genetic markers associated with inflammatory responses during mastitis could aid in the selection of diseased cattle. One potential marker is CXCR2, a chemokine receptor required for neutrophil migration to infection sites. The objective of this experiment was to identify genetic polymorphism of CXCR2 gene and associate it with subclinical and clinical mastitis. Ninety five Vrindavani crossbred cows (42-mastitis tolerant and 53-clinical mastitis) that completed at least two full lactations were taken for study. Blood of selected crossbred cows was collected, and genomic DNA was isolated by phenol chloroform method. The DNA of good quality having OD ratio (260/280 nm) between 1.7-1.9 were used for further analysis. PCR-SSCP technique was used to reveal the polymorphism in 269bp fragments of CXCR2 gene. The 269 bp fragment of CXCR2 gene was found to be monomorphic in all the DNA samples of crossbred cows

Effect of heat shock protein 70 polymorphism on thermotolerance in Tharparkar cattle

Aim: Out of various members of heat shock protein (HSP) superfamily which act a molecular chaperon by binding to the denaturing protein thus stabilizing them and preserving their activity, HSP70 are of major importance in thermotolerance development. Thus, present investigation aimed at a screening of HSP70 gene for polymorphisms and possible differences in thermotolerance in Tharparkar breed of cattle. Materials and Methods: A 295 bp fragment of HSP70 gene was subjected to polymerase chain reaction-singlestrand conformation polymorphism (SSCP) followed by sequencing of different SSCP patterns in 64 Tharparkar cattle. A comparative thermotolerance of identified genotypes was analyzed using heat tolerance coefficients (HTCs) of animals for different seasons. Results: Three SSCP patterns and consequently two alleles namely A and B were documented in one fragment of HSP70 gene. On sequencing, one single-nucleotide polymorphism with G > T substitution was found at a position that led to a change of amino acid aspartate to tyrosine in allele A. It was found that in maintaining near normal average rectal temperature, genotype AA was superior (p≤0.01). Genotype AA, thus, was found to be most thermotolerant genotype with the highest HTC (p≤0.01). Conclusion: The polymorphism at HSP70 is expected to be a potent determinant for heat tolerance in cattle, which may aid in selection for thermotolerance in cattle